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目的探讨外源性一氧化氮(NO)对日本血吸虫尾蚴的体外杀灭效果及NO抑制剂对这种杀灭作用的阻断效果。方法收集日本血吸虫感染性钉螺体内尾蚴,用RPMI 1640培养液配制成浓度为1 000条/ml的尾蚴悬液。向尾蚴悬液中加入不同浓度的NO发生剂亚硝基铁氰化钠(SNP),同时设不加SNP的对照组,观察外源性NO对尾蚴的杀灭作用及量效关系。向尾蚴悬液中加入2.00 mmol/L SNP,并加入4种NO抑制剂血红蛋白(Hb)、L半胱氨酸(L cyst)、左旋精氨酸(L arg)和硫酸亚铁(FeSO4)及其不同组合,同时设2.00 mmol/L SNP阳性对照组,观察抑制剂及其不同组合对NO杀灭尾蚴作用的阻断效应。结果经0.06、0.10 mmol/L和0.20 mmol/L SNP作用后,日本血吸虫尾蚴死亡率分别为(8.3±1.1)%、(6.26±2.3)%和(9.3±1.0)%,与对照组尾蚴死亡率差异无统计学意义(P>0.05);经0.50、1.00 mmol/L和2.00 mmol/L SNP作用后,尾蚴死亡率分别为(23.5±3.9)%、(46.0±1.1)%和(59.4±0.5)%,均显著高于对照组的尾蚴死亡率(P<0.05)。在加入2.00mmol/L SNP的同时分别加入Hb、FeSO4、L cyst、L arg、FeSO4+L cyst、FeSO4+L arg、L arg+L cyst,日本血吸虫尾蚴死亡率分别为(30.1±1.2)%、(45.1±1.4)%、(31.1±1.3)%、(34.2±3.1)%、(47.8±2.0)%、(49.1±0.6)%、(44.2±0.1)%,与2.00 mmol/L SNP阳性对照组比较,尾蚴死亡率均显著下降(P<0.05)。结论外源性NO对日本血吸虫尾蚴具有体外杀灭作用,NO抑制剂Hb、FeSO4、L cyst、L arg及其不同组合对这种杀灭作用均具有不同程度的抑制效应。
Objective To investigate the in vitro killing effect of exogenous nitric oxide (NO) on cercariae of Schistosoma japonicum and the blocking effect of NO inhibitor on this killing effect. Methods Cercariae of Schistosoma japonicum were collected and cultured in RPMI 1640 medium to make a suspension of cercariae with a concentration of 1000 / ml. To different concentrations of nitric oxide sodium cyanide (SNP) was added to the suspension of cercariae, and the control group without SNP was added to observe the killing effect of exogenous NO on cercariae and the dose-response relationship. 2.00 mmol / L SNP was added to the suspension of cercariae, and 4 NO inhibitors such as hemoglobin (Hb), L cyst, L arg and FeSO4 and The different combinations, at the same time set 2.00 mmol / L SNP positive control group, observed inhibitors and their different combinations of NO kill cercariae effect blocking effect. Results The mortality rates of cercariae of Schistosoma japonicum were (8.3 ± 1.1)%, (6.26 ± 2.3)% and (9.3 ± 1.0)%, respectively, after being treated with 0.06, 0.10 mmol / L and 0.20 mmol / The mortality of cercariae were (23.5 ± 3.9)%, (46.0 ± 1.1)% and (59.4 ±)% respectively after treated with 0.50, 1.00 mmol / L and 2.00 mmol / L SNP 0.5)%, which were significantly higher than that of the control group (P <0.05). The death rates of Schistosoma japonicum cercariae were (30.1 ± 1.2)%, respectively, when Hb, FeSO4, L cyst, L arg, FeSO4 + L cyst, FeSO4 + L arg and L arg + L cyst were respectively added with 2.00 mmol / , (45.1 ± 1.4)%, (31.1 ± 1.3)%, (34.2 ± 3.1)%, (47.8 ± 2.0)%, (49.1 ± 0.6)% and (44.2 ± 0.1)% Compared with control group, mortality of cercariae decreased significantly (P <0.05). Conclusion Exogenous NO can kill cercariae of Schistosoma japonicum in vitro. NO inhibitors Hb, FeSO4, L cyst, L arg and their different combinations have different inhibitory effects on this killing effect.